Devices, systems, and methods for removal of soluble gases from fluid samples

ABSTRACT

Devices, systems and methods are disclosed which relate to using containers with a multitude of nucleation sites covering a major portion of the inside wall of the container to enable rapid and nearly complete removal of soluble gases from fluid samples, including carbonated beverages and other carbonated fluid samples. A fluid sample is rapidly poured into the described container initiating a catastrophic release of the soluble gas from the sample.

This application claims priority to U.S. Provisional Patent ApplicationSer. No. 62/488,693, filed on Apr. 21, 2017, the contents of which arehereby incorporated by reference herein in their entirety into thisdisclosure.

TECHNICAL FIELD

The present subject disclosure relates generally to the fields ofbeverage quality control, food safety, and other applications whereefficient removal of carbon dioxide from beverages or other aqueoussamples is desirable prior to performing analytical methods, includingmethods for concentration and detection of spoilage organisms and othermicroorganisms in carbonated beverages.

BACKGROUND OF THE SUBJECT DISCLOSURE

Spoilage organisms can grow in carbonated beverages, such as widelyproduced beers, ales, and soft drinks. These organisms, while typicallyinitially present only at very low concentrations, can reproduce overtime, producing off-flavors, increased turbidity, and other qualitydefects. Further, in some cases these organisms can create safetyconcerns due to the potential for increased carbon dioxide pressure andrisk of bursting bottles.

Current methods for detection of these organisms are time-consuming andtedious, can impact the viability of entrained contaminant organisms,and are difficult to perform in an aseptic manner—thus increasing thepotential for producing erroneous results due to introducedcontaminates. Established methods include, but are not limited to,ultrasonication, filtration, combinations of ultrasonication andfiltration, automated rotary shakers, compressed air sparging (purging),and manual degassing by pouring back and forth (Smith & Marinelli,1991). The American Society of Brewing Chemists (ASBC) compared ASBCMethod Beer-1A (shaking in a flask until no further gas escapes) withgas purging, mechanical shakers, gas-permeable membrane techniques,ultrasonication, and bench-top rotary shakers (Constant & Collier,2017). The bench-top rotary shaker with baffled Erlenmeyer flasks wasdetermined to be the preferred method.

There is a great economic need for rapid identification of spoilageorganisms in carbonated beverages; both to reduce hold times prior torelease of product and to reduce the possibility of damage to thereputation and brand of the producer. Current methods of determining thepresence of contaminating microorganisms involves plating on Petridishes for identification and enumeration, or other methods of classicalmicrobiology. Rapid methods for microbiological analysis (RMMs) such aspolymerase chain reaction (PCR), immunoassays, and flow cytometry arebecoming widespread. However, use of RMMs is limited in theirapplication to the beverage industry due to the very small volumeprocessed; typically 5 to 1000 microliters. This small volume is notadequately representative of the volume of fluid in even one singlebeverage container, which is typically 200 to 1000 milliliters, up to athousand-fold greater volume than the amount analyzed.

Various conventional methods of decarbonation of beer are brieflydescribed below.

Shaking for Decarbonation: One published method of decarbonation of beersamples involves placing a beer sample into a large Erlenmeyer flask andshaking gently at first and then vigorously. While this method removescarbon dioxide and is relatively simple, it is difficult to implement inlarge laboratories performing analysis of many samples. According toPaul Smith et. al., the required analyst time for preparing 10 samplesusing this method is 25 minutes and it requires significant space due tothe large size of the flasks (500 mL flask for 200 mL of beer). SeeSmith, P. and Marinelli, L., Evaluation of Established Methods ofDecarbonating Beer. ASBC Journal. Mar. 27, 1992.

Pouring for Decarbonation: Another method for decarbonation entailspouring beer samples back and forth between beakers. This method removescarbon dioxide but is similar to the shaking method discussed above interms of labor requirements—also requiring 25 minutes to prepare 10samples. Further, this method is inherently not aseptic due to thesignificant number of times that the sample must be poured, and thus isnot a good method for use prior to spoilage organism testing.

Filtration for Decarbonation: In some instances, filtration of beersamples has been used to remove carbon dioxide. Movement of the samplefrom atmospheric pressure through a small pore and into a low pressureenvironment causes carbon dioxide to be released both on the retentateand the permeate sides of the filter. It is inherent in this processthat smaller pore sizes cause release of greater quantities of carbondioxide. This fact, in general, eliminates this method from use as afront end to the Concentrating Pipette and other membrane filtrationmethods for concentration of microorganisms, due to the need for a verylarge percentage of the target microorganisms to be present in thesample prior to and after concentration. Use of a pore size large enoughto allow larger target microorganisms to pass inherently leaves too muchcarbon dioxide in the sample, and therefor will not allow sufficientvolume to be processed with the Concentrating Pipette.

Ultrasonication for Decarbonation: Ultrasonication is capable by itselfor in combination with other methods of removing carbon dioxide frombeer samples, but it is generally not readily capable of removing carbondioxide to the levels necessary prior to processing with theConcentrating Pipette or other membrane filters. Further,ultrasonication is known to have detrimental effects on the viability ofcertain microorganisms.

Beer Glasses with Nucleation Sites. Beer glasses are now commerciallyavailable with laser cut nucleation sites on the inside wall of thebottom of the glass. These nucleation sites are used to enhance aroma,taste and head retention in beer. The nucleation surface area ispurposely limited to a very small percentage of the inside glass surfaceto create a very slow steady release of carbon dioxide in order tocreate bubbles and maintain a beer head. The nucleated surface area istypically way less than 1% of the surface area, and more often less than0.1% of the surface area. In this way, only a very small percentage ofthe contained carbon dioxide is released from the beer.

SUMMARY OF THE SUBJECT DISCLOSURE

The ability to concentrate any organisms present in a significant amountof the beverage will increase the utility of RMMs by increasing thelikelihood of early detection, benefiting industry and consumers.Applicant specializes in membrane-based concentration of biologicalparticles from fluids and has been awarded six US patents in this areato-date; U.S. Pat. Nos. 8,110,112, 8,758,623, 8,584,535, 9,593,359,9,574,977, and 8,726,744, all of which are incorporated by referenceherein in their entirety into this disclosure. Three commercialbiological concentrators have been developed and sold, based on thepatented process, termed “WET FOAM ELUTION”, the HSC-40, HCl-40, andCP-150 Concentrating Pipette (current data sheets available atwww.innovaprep.com).

As used herein and throughout this disclosure, the Concentrating Pipetteis as described in the various patents incorporated by reference in thisdisclosure. An example of such a Concentrating Pipette Instrument isshown in FIG. 10. One of the advantages of the present subjectdisclosure is in preparing a sample size from a carbonated fluid to theConcentrating Pipette device.

While Applicant has demonstrated that the concentration of biologicalparticles from a carbonated fluid can be performed using the patentedmethods and commercial instruments at increased pressure, thus keepingthe gas in solution, Applicant's patented method of concentration wasnot effective for carbonated beverage samples at ambient pressures. Areason for this is because the vacuum applied to the filter releases gasfrom the fluid as it is drawn through the membrane, blocking the poresin the membrane. The solubility of carbon dioxide in water (and thus analcoholic beverage, e.g., beer) at various temperatures is significant.FIG. 1 shows high solubility of carbon dioxide in water, particularly atlower temperatures.

In the present subject disclosure, Applicants have demonstrated a novelmethod for rapid degassing of carbonated beverage samples by pouringthem into an etched or “frosted” glass or plastic container with enoughvolume to hold the sample as it catastrophically foams up, evolves themajority of the carbon dioxide gas, and collapses as flat liquid in thebottom of the container.

This process uses a container that has been treated using sandblastingor other methods to create a multitude of nucleation sites on a largesurface area on the inside of the container. By creating nucleationsites over a major portion of the inside area of the container, thenucleation sites not only cause a release of carbon dioxide, but alsoinitiate a catastrophic release of carbon dioxide through formation of amultitude of gas bubbles causing significant mixing and additionalrelease of gas.

This catastrophic release of carbon dioxide is efficient enough thatafter several minutes of settling, enough carbon dioxide is evolved thatthe sample is free enough of carbon dioxide to allow for processingthrough a membrane filter such as those in the Concentrating PipetteTips used in the Concentrating Pipette instrument.

Further, vigorous pouring of the sample into the container furtherenhances the catastrophic release of carbon dioxide and further reducesthe amount of residual carbon dioxide present. In some instances, theuse of a mild heating step prior to pouring into the container can beused to reduce the solubility of carbon dioxide in the sample to furtherenhance the release of carbon dioxide from the sample. Finally,refrigeration of the sample, immediately after pouring into thedecarbonation container or several minutes later, can further enhancethe ability to process through membrane filters by increasing thesolubility of the carbon dioxide—making it more difficult to removeduring the membrane filtration process.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated in and constitute apart of this specification, illustrate embodiments of the subjectdisclosure and technical data supporting those embodiments, and togetherwith the written description, serve to explain certain principles of thesubject disclosure.

FIG. 1 shows the solubility of carbo dioxide in water.

FIG. 2 shows a decarbonation container, according to an exemplaryembodiment of the present subject disclosure.

FIG. 3 shows a vented lid for a decarbonation container, according to anexemplary embodiment of the present subject disclosure.

FIG. 4 shows a nucleated PYREX measuring cup, according to an exemplaryembodiment of the present subject disclosure.

FIG. 5 shows beer poured into a nucleated container, according to anexemplary embodiment of the present subject disclosure.

FIG. 6 shows flattened beer being concentrated, according to anexemplary embodiment of the present subject disclosure.

FIG. 7 shows a 12 ounce can of flattened beer filtered, according to anexemplary embodiment of the present subject disclosure.

FIG. 8 shows nucleation sites being created inside a container,according to an exemplary embodiment of the present subject disclosure.

FIG. 9 shows an internally nucleated container, according to anexemplary embodiment of the present subject disclosure.

FIG. 10 shows a Concentrating Pipette Instrument, according to anexemplary embodiment of the present subject disclosure.

DETAILED DESCRIPTION OF THE SUBJECT DISCLOSURE

The following detailed description references specific embodiments ofthe subject disclosure and accompanying figures, including therespective best modes for carrying out each embodiment. It shall beunderstood that these illustrations are by way of example and not by wayof limitation.

The present subject disclosure describes highly efficient and simple touse devices, systems, and methods for removing saturated gasses, such ascarbon dioxide, from liquid samples. The technique uses a large surfacearea of nucleation sites along with methods for enhancing the nucleationof the gasses to quickly and efficiently remove these gases from samplesprior to implementing liquid concentration or other laboratory methods.Specifically, the present technique may be used to decarbonate beer andother carbonated liquids prior to processing on a Concentrating PipetteInstrument, for example those described and patented by the Applicant.

FIG. 2 shows a decarbonation container 100, according to an exemplaryembodiment of the present subject disclosure. A large zone of nucleationsites 101, on the inside surface of the container, extends from near theneck at the top end to the bottom edge of the inside walls and includesthe entire bottom inside surface as well. Thus, any fluid which ispoured into container 100 will come into direct contact with thenucleation sites 101, which extend to nearly the entire interior surfaceof the container 100. The higher the percentage of inside walls ofcontainer 100 is covered by nucleation sites 101, the more effective thedecarbonation of a fluid deposited therein. Typically, the nucleatedinterior surface is 5%-100% of the interior surface. Further, a standardbottle thread 102 is provided to allow for use of a threaded lid.

FIG. 3 shows a screw top lid 200 including a hydrophobic membrane filtervent 201 for use on container 100. The user pours a carbonated sampleinto container 100 and carbon dioxide is nucleated by nucleation surface101. After pouring the sample into the container 100, the user may placelid 200 onto container 100 to reduce the potential for contamination ofthe liquid sample. Vent 201 allows gas to escape during thedecarbonation process so that pressure does not build up withincontainer 100.

Applicant has demonstrated sufficient rapid degassing of carbonatedbeverage samples by pouring them into glass or plastic containers thathave had a major portion of the inside walls etched or “frosted” andwhich contain enough volume to hold the sample as it catastrophicallyfoams up, evolving the majority of the carbon dioxide gas, andcollapsing as flat liquid in the bottom of the container.

An exemplary demonstration of the process was accomplished by gritblasting a 2 Liter Pyrex Measuring Cup (see FIG. 4). The blastingprocess “frosted” the interior of the container, forming nucleationsites over the internal surface. When a can or bottle of beer was pouredinto the container, it immediately foamed up to the top, then rapidlycollapsed back to a flat liquid with very little residual carbonation(FIG. 5). As is shown in FIGS. 6-7, the entire bottle or can is thenable to be rapidly processed using the Concentrating Pipette Instrument.In the example shown, the CP-150 Concentrating Pipette Instrument wasused. This Instrument is shown again in FIG. 10. As an example, a 12ounce can of Busch Light beer was filtered in 7 minutes and 43 seconds.

Various other glass and plastic containers may be treated by interiorgrit blasting and have demonstrated to behave in the same manner. Such agrit blasting process is shown in FIG. 8. Various beers and ales,including but not limited to Angry Orchard hard cider, Boulevardunfiltered wheat, single wide IPA, Tank 7 Farmhouse Ale, and Pale Ale;Budweiser, Carlsberg, Coors Banquet, Miller Lite, Guinness Draught,Heineken, Modelo Especial, Samuel Adams Boston Lager, Sapporo, ShinerBock, Stella Artois, and TsingTao were shown to be quickly decarbonatedusing the nucleated containers.

Nucleation sites may be created on the container in a number of waysincluding, but not limited to: by use of a mold during manufacturingthat contains a reverse image of the nucleation sites, by use of acontainer material that is naturally rough or naturally contains manynucleation sites, or by post treatment of the container after initialmanufacturing using any number of well-known processes for creating arough surface. These processes include: sandblasting, bead blasting,laser etching, acid etching, or other mechanical or chemical means forroughening the surface or creating nucleation sites.

Molding may also be used to create a rough surface, wherein thenucleation sites are produced during molding of the container, by usinga mold containing a mirror image of the nucleation sites.

Sandblasting may be performed using aluminum oxide, silicon carbide,crushed glass, glass beads, plastic abrasive, pumice, steel shot, steelgrit, corn cob, walnut shell or garnet with a grit ranging from 8 to1,000. Laser etching may be performed with any number of commerciallyavailable laser cutter/etcher systems. Plastic containers, especially,may be manufactured with a multitude of nucleation sites by using a moldthat has be roughened using sandblasting or other methods.

The nucleation sites may be created in any number of types of materialsincluding, but not limited to: glass, ceramic, metal, plastic,thermoplastic, polytetrafluoroethylene or other material routinely usedfor producing bottles or sample tubes for laboratory use. The containermay be autoclavable, such that it can be washed and then autoclavedbetween samples to ensure that no cross contamination occurs betweensamples. Alternatively, the container may be single use and may bepackaged and treated, prior to sale, using e-beam irradiation, gammairradiation, ethylene oxide, vaporous hydrogen peroxide, peracetic acidor other commonly used sterilization processes.

The container is sized such that entire sample may be vigorously pouredinto the container and not overflow due to the decarbonation process andassociated foaming of the sample. In the case of beer, most sample 350mL cans or bottles of beer can be rapidly poured into a container of 1 Lnominal volume without overflowing during the decarbonation process.Other samples types and volumes may require larger containers or may beable to be processed in smaller containers.

Applicants have proven the effectiveness of the present technique withvarious volumes. For example, from 100 mL of beer to 750 mL of sparklingwine. Degassing of up to 12 ounces (335 mL) of beer has been performedin 32 ounce straight sided jars (like the jars shown in FIGS. 2 and 9).A 750 mL wine sample was degassed in a 60 ounce sand blasted beerpitcher and was run on the Concentrating Pipette with a 0.4 umConcentrating Pipette Tip in 12 minutes and 35 seconds. The presenttechnique is effective for a wide range of volumes, from 5 mL to 10 L.Further, different shapes of containers may also be used to enhance thedecarbonation process through increased turbulence associated withdifferent container diameters and shapes.

A large surface area with nucleation sites may also be produced by usinga multitude of devices such as glass beads or a single device or severaldevices with a large surface area of nucleation sites which are placedinto the container prior to pouring the sample.

The decarbonation process causes significant quantities of gas to bereleased and as such the process to some extent helps maintain sterilitydue to an outflow of carbon dioxide from the container. Further thisoutflow requires that an open container or some type of vent be usedduring the decarbonation process. For example, a loose-fitting lid maybe used to eliminate the chance of a user touching the container openingwhile also constricting the open area enough to ensure only an outflowof carbon dioxide takes place. Further, lids with an integral membranefilter of a small enough pore size to ensure sterility may also be usedto allow for an outflow of gas while helping to reducing the chance ofcontamination.

In addition to the decarbonation action performed by use of a samplecontainer with a large surface area of nucleation sites, other steps maybe taken to enhance the rate and completeness of the decarbonationprocess. These include vigorous pouring of the beer into the container,heating of the sample prior to pouring into the container, stirring orshaking of the sample while in the container, as well as other methodsfor enhancing mixing and contact with the nucleation sites. Further, theability to process the sample following decarbonation may also beenhanced by cooling the sample during or after the decarbonation processto increase the solubility of the remaining carbon dioxide in the sampleand thus make it more difficult to come out of solution duringsubsequent filtration or concentration processes.

Vigorous pouring of beer samples or other carbonated or gassed samplesmay entail turning the sample container—in the case of beer the bottleor can—nearly or completely upside down from a height of a few inches toone foot or more in height above the nucleated container and letting theentire sample flow into the container.

Heating of the sample includes any temperature from room temperature toup to 37° C. or more to enhance the release of carbon dioxide. In thecase of beer tests were performed by putting full, unopened cans orbottles of beer in a 37° C. incubator for 30 minutes before pouring intothe container. This significantly enhanced the carbon dioxide removaland reduced the total time period required for decarbonation and sampleprocessing on the Concentrating Pipette.

Further, in addition to beer, other sample types such as cider,sparkling wine, champagne, wine coolers, other alcoholic beverages,juice, lemonade, coffee, soft drink, coke, fizzy drink, fizzy juice,cool drink, cold drink, lolly water, pop, seltzer, soda, soda pop,fountain drink, ginger ale, ginger beer, tonic water, mineral water, orother carbonated beverages may also be decarbonated using this method.Additionally, other fluid samples containing carbon dioxide, nitrogen,nitrous oxide, oxygen or other soluble gases or mixtures of solublegases may also be degassed using these methods.

The more complete the removal of carbon dioxide from the beer orcarbonated beverage the more quickly and completely the sample may beprocessed on the Concentrating Pipette. Without decarbonation of beersamples, most beer fouls the Concentrating Pipette Tip and causes theinstrument to shut down within seconds. Provided below in Table 1through Table 7 are data for processing of a number of brands and stylesof beer following decarbonation. Table 1 through Table 5 show varyingvolumes of beer processed and varying run times for different types ofbeers and different types of Concentrating Pipette Tips. Table 6 andTable 7 provide a comparison between beer samples at room temperatureand beer samples heated at 37° C. prior to processing.

Membrane filtration and concentration of biological particles includingbeer spoilage organisms spiked into Coors beer was then demonstratedusing the Concentrating Pipette equipped with 0.45 micron hollow fiberpipette tips, P/N CC 08018. The first three test runs (test runs 1-3)are shown with a prototype next generation instrument with 3.5 psibackpressure and demonstrate an average of 693X concentration of theorganism in an average of 3.6 minutes (see Table 6). Test runs 4-6 usedthe current generation instrument (CP-150) and test runs 7-9 used thenext generation instrument (CP SELECT) without backpressure, with thedifference being that the next generation instrument has improvedvalves, foam control, software, etc.

Optimization of the grit blasting (nucleating) process (FIG. 8) wasundertaken using a commercially available autoclavable culture vessel(32 oz wide mouth glass jar with HEPA-vent lid, P/N C607, PhytoTechnology Laboratories, Mission, Kans. USA). The vented lid allows thegas to escape during decarbonation without building up pressure in thejar. Aluminum Oxide sandblasting media and other media were screened forthe ability to create an optimum surface for beer decarbonation,including but not limited to 150 grit blasted for 8 minutes which wasselected for production of decarbonation vessels for commercial sale.Other values and time ranges are also possible and within the purview ofthe present disclosure.

The blasting media size may range from 8-1,000 grit, and morespecifically would be limited to 18-500 grit. Above 500 grit is the“micro fine” area and less conducive to creation of nucleation sites.Testing times have ranged from 1 minute (minimum to lightly blast thejar) to 32 minutes. The 32 minutes involved several instances ofsandblasting with a fine grit blasting media. As such, the process wasto blast, test, and repeat. Beyond 30 minutes is where effectivenessstarts to drop. An exemplary preferable material used is 150 gritaluminum oxide for 6-8 minutes at ˜100 psi. An internally nucleated jaris shown in FIG. 9.

In another exemplary embodiment, Styrofoam cups and containers are used.Styrofoam cups and containers inherently provide a vast number ofnucleation sites due to their method of manufacture. Observation of suchcups under a microscope clearly shows nucleation sites where cellsborder each other and where edges are created on the cells in theinterstices between cells by injection into mold cavities.

Three samples were degassed in Styrofoam cups and processed on theConcentrating Pipette Instrument (CP-150, FIG. 10). The appearance ofthe degassing process was similar to that in a frosted glass container.After pouring the beer in the cups, the standard lids with either theslit aperture for inserting a drinking straw, or the coffee drinking lidwith the small sipping hole, were applied to the cups. Due to gasevolution from the cups, these small gas openings are sufficient toallow the evolving gas to escape without blowing the lids off. Theevolving gas may also protect the sample from contamination during thesettling and degassing step. In this case, the samples were chilled foran hour or less. See Table 9 for results of beer degassing usingcommercially available Styrofoam cups.

Following the above exemplary treatments, the Applicant ConcentratingPipette and other Applicant instruments based on the above patents andpending patent application Ser. Nos. 14/313,618, 15/456,981, 15/431,655,and 14/058,193 are able to concentrate the initial sample into a volumeof from less than 100 to 1000 milliliters or more in a matter ofminutes.

It will be appreciated that the foregoing instrumentalities teach by wayof example, and not by limitation. Accordingly, those skilled in the artunderstand that the subject matter is not limited to what is strictlydisclosed, but also pertains to what is understood by those skilled inthe art on the basis of the teachings herein. The inventors hereby statetheir subject matter to rely, as may be needed, upon the Doctrine ofEquivalents to protect the fullness of their rights in what is claimed.

TABLE 1 Concentration Pipette Run Times for Decarbonated Beer - BestResults (with 4° C. chilling) Average Time at Volume time 4° C. Can/Number Beer Tip processed (min) (min) bottle of runs Coors 0.4 um all13.86 20 (as both 16 Banquet flat (355 mL) low as 3) Heinekin 0.4 um all17.26 20 Bottle 7 flat (355 mL) Stella 0.4 um all 8.82 20 Bottle 3Artoise flat (355 mL) TsingTao 0.45 um All 8.7 20 Bottle 3 hf-CPT (355mL)

TABLE 2 Concentrating Pipette Run Times for Decarbonated Beer - 2^(nd)Best Results (with 4° C. chilling) Average Average Time at volume time4° C. Can/ Number Beer Tip processed (min) (min) bottle of runsBudweiser 0.4 um 265.33 mL 22.74 20 Can 3 flat Shiner 0.4 um  259.9 mL34.13 20 Bottle 2 Bock flat Modelo 0.4 um 167.67 mL 10.96 20 Can 3Especial flat Guinness 0.45 um 170.11 16.74 20 Both 4 Draught hf-CPTSapporo 0.45 um 238.99 17.5 20 Can 1 hf-CPT

TABLE 3 Concentrating Pipette Run Times for Decarbonated Beer - 3^(rd)Best Results (with 4° C. chilling) Avg volume Average Time at processedtime 4° C. Can/ Number Beer Tip (mL) (min) (min) bottle of runsBoulevard 0.4 um 147.68 10.81 20 Bottle 4 Single Wide flat IPA

TABLE 4 Concentrating Pipette Run Times for Decarbonated Beer - 4^(th)Best Results (with 4° C. chilling) Avg volume Average Time at processedtime 4° C. Can/ Number Beer Tip (mL) (min) (min) bottle of runs Samuel0.4 um 153.83 29.79 20 Can 2 Adams flat Boston Lager Angry 0.4 um 127.7031.44 20 Bottle 2 Orchard had flat cider Sapporo 0.4 um 127.27 19.33 20Can 1 flat Boulevard 0.45 um 147.68 11.1 20 bottle 2 Pale Ale hf-CPT

TABLE 5 Concentrating Pipette Run Times for Decarbonated Beer - 5^(th)Best Results (with 4° C. chilling) Average Average Time at volume time4° C. Can/ Number Beer Tip processed (min) (min) bottle of runsCarlsberg 0.4 um 32.54 31.83 20 Can 2 flat Boulevard 0.4 um 78.40 11.7120 Bottle 2 Pale Ale flat Boulevard 0.4 um 32.64 27.43 20 Bottle 1 Tank7 flat Boulevard 0.45 um 109.11 27.58 20 Bottle 1 Tank 7 hf-CPTBoulevard 0.45 um 63.72 11.72 20 Bottle 1 unfiltered hf-CPT wheat

TABLE 6 Concentration Pipette Run Times for Decarbonated Beer -Comparison −25° C. and 37° C. prior to decarbonation) (with 4° C.chilling) Temp Average Average prior to volume time Can/ Number Beerpouring processed (min) Tip type bottle of runs Coors 25 355 13.86 0.4um Can 16 Banquet flat Coors 37 355 6.53 0.4 um Can 3 Banquet flatBudweiser 25 255.33 22.74 0.4 um Can 3 flat Budweiser 37 452.76 16.510.4 um Can 3 flat Heinekin 25 354.1 17.26 0.4 um Bottle 7 flat Heinekin37 349.82 10.8 0.4 um Bottle 1 flat

TABLE 7 Concentration Pipette Run Times for Decarbonated Beer -Comparison −25 ° C. and 37° C. prior to decarbonation) (with 4° C.chilling) Temp Average Average prior to volume time Can/ Number Beerpouring processed (min) Tip type bottle of runs TsingTao 25 119.74 20.910.4 um Bottle 3 flat TsingTao 37 131.98 6.27 0.4 um Bottle 1 flat StellaArtois 25 328.53 8.81 0.4 um Bottle 3 flat Stella Artois 37 324.96 6.220.4 um Bottle 1 flat

TABLE 8 InnovaPrep Concentration of Beer Spoilage Organism fromDecarbonated Beer L. brevis spiked Miller High Life Mar. 14, 2017 1 2 3avg st dev 4 5 6 Runs time 3.00 4.00 3.82 3.6067 0.4352 6.70 4.82 4.17Feed Titer (~1 CFU/mL) 196 196 plated 100 uL/counts 187 187 164 164average 182.3333 182.3333 dilution 1.0000 1.0000 total spike 182.3333182.3333 feed/titer, CFU/mL 0.5136 0.5136 Concentrate tare 3.7157 3.71493.69 3.7033 3.7036 3.7017 net 3.9074 4.0485 4.0881 4.3726 4.4142 4.5058volume 0.1917 0.3336 0.3981 0.3078 0.0862 0.6693 0.7106 0.8041 counts,CFUs 93 99 114 106 105 102 conctiter CFU/mL 485.1330 296.7626 286.3602158.3744 147.7625 126.8499 % Efficiency 51.01% 54.30% 62.52% 55.94%4.84% 58.14% 57.59% 55.94% concentration factor for 944.5460 577.7919557.5386 693.2922 177.8556 308.3524 287.6911 246.9747 Extraction 2 tare3.6966 3.7267 3.6951 3.6965 3.6936 3.716 net 3.9142 4.0673 3.9811 4.39054.4625 4.4615 volume 0.2176 0.3406 0.286 0.2814 0.0503 0.6940 0.76890.7455 counts, CFU 36 9 9 22 4 3 Efficiency 19.74% 4.94% 4.94% 9.87%6.98% 12.07% 2.19% 1.65% total elution volume 0.4093 0.6742 0.68411.3633 1.4795 1.5496 total efficiency 70.75% 59.23% 67.46% 65.81% 4.84%70.20% 59.78% 57.59% avg st dev 7 8 9 avg st dev Runs time 5.2300 1.07284.13 3.75 3.73 3.8700 0.1840 Feed Titer (~1 CFU/mL) 196 plated 100uL/counts 187 164 average 182.3333 dilution 1.0000 total spike 182.3333feed/titer, CFU/mL 0.5136 Concentrate tare 3.7258 3.7249 3.688 net4.1903 4.2205 4.0338 volume 0.7280 0.0564 0.4645 0.4956 0.3458 0.43530.0645 counts, CFUs 117 111 81 conctiter CFU/mL 251.8837 223.9709234.2394 % Efficiency 57.22% 0.93% 64.17% 60.88% 44.42% 56.49% 8.64%concentration factor for 281.0060 25.4993 490.4135 436.0677 456.0603460.8472 22.4433 Extraction 2 tare 3.6991 3.7065 3.6897 net 4.17504.1963 4.086 volume 0.7361 0.0313 0.4759 0.4898 0.3963 0.4540 0.0412counts, CFU 10 6 2 Efficiency 5.30% 4.79% 5.48% 3.29% 1.10% 3.29% 1.79%total elution volume 0.9404 0.9854 0.7421 total efficiency 62.52% 5.50%69.65% 64.17% 45.52% 59.78% 10.33%

TABLE 9 Beer Degassing Using Commercially Available Styrofoam Cups Proc-Proc- essed Chill essing Sam- 1st Sam- time Time ple Elution ple BeerDegassing Vessel (min) (m:s) (g) (uL) 1 Miller 32 Oz. Styrofoam 40 7:03277 380 High Life Cup (Dart 32AJ20) 2 Coors 32 Oz. Styrofoam 50 6:24 266260 Banquet Cup (Dart 32AJ20) 3 Miller 12 Oz. Styrofoam 60 3:38 118 350High Life (Sweetheart X12 67240)

What is claimed is:
 1. A system for degassing a fluid sample, the systemcomprising a container with a plurality of nucleation sites for exposingto the fluid sample, wherein a total area of the nucleation sites is atleast 5% of a total surface area of contact of the fluid sample with thecontainer, and wherein a lid with open holes or holes covered with amembrane filter material is used to allow gas to escape duringdegassing.
 2. The system of claim 1, wherein the fluid sample containscarbon dioxide, nitrogen, nitrous oxide, oxygen or other soluble gasesor mixtures of soluble gases.
 3. The system of claim 1, wherein thefluid sample is one of a carbonated beverage, beer, cider, sparklingwine, champagne, wine cooler, alcoholic beverage, juice, lemonade,coffee, soft drink, coke, fizzy drink, fizzy juice, cool drink, colddrink, lolly water, pop, seltzer, soda, soda pop, fountain drink, gingerale, ginger beer, tonic water, mineral water, or other carbonatedbeverage.
 4. The system of claim 1, wherein the container is made fromone of glass, Styrofoam, ceramic, metal, plastic, thermoplastic,polytetrafluoroethylene or other material routinely used for producingbottles or sample tubes for laboratory use.
 5. The system of claim 1,wherein the nucleation sites are produced during molding of thecontainer, by using a mold containing a mirror image of the nucleationsites.
 6. The system of claim 1, where in the nucleation sites areproduced after molding of the container by sandblasting, laser etching,machining, acid etching or other chemical or mechanical means ofcreating nucleation sites on the surface.
 7. The system of claim 6,wherein the sandblasting is performed using aluminum oxide, siliconcarbide, crushed glass, glass beads, plastic abrasive, pumice, steelshot, steel grit, corn cob, walnut shell or garnet with a grit rangingfrom 8 to 1,000.
 8. A method for degassing a fluid, the methodcomprising: pouring a fluid sample into a container with a plurality ofnucleation sites, wherein a total area of the nucleation sites is atleast 5% of a total surface area of contact of the fluid sample with thecontainer and wherein a lid with open holes or holes covered with amembrane filter material is used to allow gas to escape duringdegassing; and incubating the fluid in the container for a sufficienttime to allow for degassing.
 9. The method of claim 8, wherein the fluidsample is poured into the container in such a manner to agitate thefluid sample and increase the rate of degassing.
 10. The method of claim8, wherein the fluid sample is first heated to increase the rate ofdegassing.
 11. The method of claim 8, wherein the fluid is cooled duringor after degassing to increase the solubility of the gas in the fluid.12. A system for degassing a fluid sample, the system comprising acontainer with a plurality of nucleation sites for exposure to the fluidsample, wherein a total area of the nucleation sites is at least 5% of atotal surface area of contact of the fluid sample with the container,wherein the nucleation sites are on a device or plurality of deviceswhich are placed in the container, and wherein a lid with open holes orholes covered with a membrane filter material is used to allow gas toescape during degassing.
 13. The system of claim 12, wherein thecontainer is made from one of glass, Styrofoam, ceramic, metal, plastic,thermoplastic, polytetrafluoroethylene or other material routinely usedfor producing bottles or sample tubes for laboratory use.
 14. The systemof claim 12, where in the nucleation sites are produced during moldingof the container, by using a mold containing a mirror image of thenucleation sites.
 15. The system of claim 12, where in the nucleationsites are produced after molding of the container by sandblasting, laseretching, machining or other mechanical means of creating nucleationsites on the surface.
 16. The system of claim 15, wherein thesandblasting is performed using aluminum oxide, silicon carbide, crushedglass, glass beads, plastic abrasive, pumice, steel shot, steel grit,corn cob, walnut shell or garnet with a grit ranging from 8 to 1,000.17. The system of claim 12, wherein the container is of sufficient sizeto allow the fluid sample to foam while not overflowing the container.